Pyrazolopyrimidines as crf receptor antagonists

ABSTRACT

This invention concerns compounds of formula  
                 
 
     including the stereoisomers and the pharmaceutically acceptable acid addition salt forms thereof, wherein R 1  is NR 4 R 5  or OR 5 ; R 2  is C 1-6 alkyl, C 1-6 alkyloxy or C 1-6 alkylthio; R 3  is hydrogen, C 1-6 alkyl, C 1-6 alkylsulfonyl, C 1-6 alkylsulfoxy or C 1-6 alkylthio; R 4  is hydrogen, C 1-6 alkyl, mono- or di(C 3-6 cycloalkyl)methyl, C 3-6 cycloalkyl, C 3-6 alkenyl, hydroxyC 1-6 alkyl, C 1-6 alkylcarbonyloxyC 1-6 alkyl or C 1-6 alkyloxyC 1-6 alkyl; R 5  is C 1-8 alkyl, mono- or di(C 3-6 cycloalkyl)methyl, Ar 1 CH 2 , C 1-6 alkyloxyC 1-6 alkyl, hydroxyC 1-6 alkyl, C 3-6 alkenyl, thienylmethyl, furanylmethyl, C 1-6 alkylthioC 1-6 alkyl, morpholinyl, mono- or di(C 1-6 alkyl)aminoC 1-6 alkyl, di(C 1-6 alkyl)amino, C 1-6 alkylcarbonylC 1-6 alkyl, C 1-6 alkyl substituted with imidazolyl; or a radical of formula —Alk-O—CO—Ar 1 ; or R 4  and R 5  taken together with the nitrogen atom to which they are attached may form an optionally substituted pyrrolidinyl, piperidinyl, homopiperidinyl or morpholinyl group; having CRF receptor antagonistic properties; pharmaceutical compositions containing such compounds as active ingredients; methods of treating disorders related to hypersecretion of CRF such as depression, anxiety, substance abuse, by administering an effective amount of a compound of formula (I).

BACKGROUND OF THE INVENTION

[0001] This invention relates to pyrazolopyrimidines which possess CRFreceptor antagonistic properties, to pharmaceutical compositionscontaining these compounds as active ingredient, and the use thereof inthe treatment of endocrine, psychiatric and neurologic conditions orillnesses, including stress-related disorders in general.

[0002] The first corticotropin-releasing factor (CRF) was isolated fromovine hypothalmi and identified as a 41-amino acid peptide (Vale et al.,Science 213:1394-1397, 1981). Subsequently, sequences of human and ratCRF were isolated and determined to be identical, but different fromovine CRF in 7 of the 41 amino acid residues (Rivier et al., Proc. Natl.Acad. Sci. USA 80:4851, 1983; Shibahara et al., EMBO J. 2:775, 1983).CRF has been found to produce profound alterations in endocrine, nervousand immune system functions. CRF is believed to be the majorphysiological regulator of the basal and stress-release ofadrenocorticotropic hormone (“ACTH”), β-endorphin, and otherpro-opiomelanocortin (“POMC”)-derived peptides from the anteriorpituitary (Vale et al., Science 213:1394-1397, 1981). Briefly, CRF isbelieved to initiate its biological effects by binding to a plasmamembrane receptor which has been found to be distributed throughout thebrain (DeSouza et al., Science 221:1449-1451, 1984), pituitary (DeSouzaet al., Methods Enzymol. 124:560, 1986; Wynn et al., Biochem. Biophys.Res. Comm. 110:602-608, 1983), adrenals (Udelsman et al., Nature319:147-150, 1986) and spleen (Webster, E. L., and E. B. DeSouza,Endocrinology 122:609-617, 1988). The CRF receptor is coupled to aGTP-binding protein (Perrin et al., Endocrinology 118: 1171-1179, 1986)which mediates CRF-stimulated increase in intracellular production ofcAMP (Bilezikjian, L. M., and W. W. Vale, Endocrinology 113:657-662,1983).

[0003] In addition to its role in stimulating the production of ACTH andPOMC, CRF is also believed to coordinate many of the endocrineautonomic, and behavioral responses to stress, and may be involved inthe pathophysiology of affective disorders. Moreover, CRF is believed tobe a key intermediary in communication between the immune, centralnervous, endocrine and cardiovascular systems (Crofford et al., J. Clin.Invest. 90:2555-2564, 1992; Sapolsky et al., Science 238:522-524, 1987;Tilders et al., Regul. Peptides 5:77-84, 1982). Overall, CRF appears tobe one of the pivotal central nervous system neurotransmitters and playsa crucial role in integrating the body's overall response to stress.

[0004] Administration of CRF directly to the brain elicits behavioral,physiological, and endocrine responses identical to those observed foran animal exposed to a stressful environment. For example,intracerebroventricular injection of CRF results in behavioralactivation (Sutton et al., Nature 297:331, 1982), persistent activationof the electroencephalogram (Ehlers et al., Brain Res. 2/8332, 1983),stimulation of the sympathoadrenomedullary pathway (Brown et al.,Endocrinology 110:928, 1982), an increase of heart rate and bloodpressure (Fisher et al., Endocrinology 110:2222, 1982), an increase inoxygen consumption (Brown et al., Life Sciences 30:207, 1982),alteration of gastrointestinal activity (Williams et al., Am. J. Physiol253:G582, 1987), suppression of food consumption (Levine et al.,Neuropharmacology 22:337, 1983), modification of sexual behavior(Sirinathsinghji et al., Nature 305:232, 1983), and immune functioncompromise (Irwin et al., Am. J. Physiol. 255:R744, 1988). Furthermore,clinical data suggest that CRF may be hypersecreted in the brain indepression, anxiety-related disorders, and anorexia nervosa. (DeSouza,Ann. Reports in Med. Chem. 25:215-223, 1990).

[0005] Accordingly, clinical data suggest that CRF receptor antagonistsmay represent novel antidepressant and/or anxiolytic drugs that may beuseful in the treatment of the neuropsychiatric disorders manifestinghypersecretion of CRF. CRF receptor antagonists have been reported infor example, U.S. Pat. No. 5,063,245 disclosing substituted4-thio-5-oxo-3-pyrazoline derivatives and Australian Patent No.AU-A-41399/93, disclosing substituted 2-aminothiazole derivatives.WO-92/18504 and JP-32/04877 disclose pyrazolo[1,5-a]pyrimidines asantiinflammatory agents. Also, WO-94/13676, WO-94/13677 and WO-95/33750disclose pyrrolopyrimidines, pyrazolo[3,4-d]pyrimidines and substitutedpurines as CRF receptor antagonists. Arylpyrazolo[1,5-a]pyrimidines havebeen described as xanthine oxidase inhibitors (Robins et al., J.Heterocyclic Chem. 22:601-634, 1985). JP-42/011,753 discloses7-methylamino-pyrazolo[1,5-a]pyrimidine derivatives useful as sedativeand antiphlogistic agents. And JP-61/057,587 disclosespyrazolo[1,5-a]pyrimidine derivatives useful as antiulcer agents.

[0006] Due to the physiological significance of CRF, the development offurther biologically active small molecules having significant CRFreceptor binding activity and which are capable of antagonizing the CRFreceptor remains a desirable goal. Such CRF receptor antagonists wouldbe useful in the treatment of endocrine, psychiatric and neurologicconditions or illnesses, including stress-related disorders in general.

DESCRIPTION OF THE INVENTION

[0007] This invention concerns CRF antagonistic compounds of formula (I)

[0008] including the stereoisomers and the pharmaceutically acceptableacid addition salt forms thereof, wherein

[0009] R¹ is NR⁴R⁵ or OR⁵;

[0010] R² is C₁₋₆alkyl, C₁₋₆alkyloxy or C₁₋₆alkylthio;

[0011] R³ is hydrogen, C₁₋₆alkyl, C₁₋₆alkylsulfonyl, C₁₋₆alkylsulfoxy orC₁₋₆alkylthio;

[0012] R⁴ is hydrogen, C₁₋₆alkyl, mono- or di(C₃₋₆cycloalkyl)methyl,C₃₋₆cycloalkyl, C₃₋₆alkenyl, hydroxyC₁₋₆alkyl,C₁₋₆alkylcarbonyloxyC₁₋₆alkyl or C₁₋₆alkyloxyC₁₋₆alkyl;

[0013] R⁵ is C₁₋₈alkyl, mono- or di(C₃₋₆cycloalkyl)methyl, Ar¹CH₂,C₁₋₆alkyloxyC₁₋₆alkyl, hydroxyC₁₋₆alkyl, C₃₋₆alkenyl, thienylmethyl,furanylmethyl, C₁₋₆alkylthioC₁₋₆alkyl, morpholinyl, mono- ordi(C₁₋₆alkyl)aminoC₁₋₆alkyl, di(C₁₋₆alkyl)amino,C₁₋₆alkylcarbonylC₁₋₆alkyl, C₁₋₆alkyl substituted with imidazolyl; or aradical of formula —Alk—O—CO—Ar¹;

[0014] or R⁴ and R⁵ taken together with the nitrogen atom to which theyare attached may form a pyrrolidinyl, piperidinyl, homopiperidinyl ormorpholinyl group, optionally substituted with C₁₋₆alkyl orC₁₋₆alkyloxyC₁₋₆alkyl; and

[0015] Ar is phenyl; phenyl substituted with 1, 2 or 3 substituentsindependently selected from halo, C₁₋₆alkyl, trifluoromethyl, hydroxy,cyano, C₁₋₆alkyloxy, benzyloxy, C₁₋₆alkylthio, nitro, amino and mono- ordi(C₁₋₆alkyl)amino; pyridinyl; pyridinyl substituted with 1, 2 or 3substituents independently selected from halo, C₁₋₆alkyl,trifluoromethyl, hydroxy, cyano, C₁₋₆alkyloxy, benzyloxy, C₁₋₆alkylthio,nitro, amino, mono- or di(C₁₋₆alkyl)amino and piperidinyl; and whereinsaid substituted phenyl may optionally be further substituted with oneor more halogens;

[0016] Ar¹ is phenyl; phenyl substituted with 1, 2 or 3 substituentseach independently selected from halo, C₁₋₆alkyl, C₁₋₆alkyloxy,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, trifluoromethyl and C₁₋₆alkyl substitutedwith morpholinyl; or pyridinyl; and

[0017] Alk is C₁₋₆alkanediyl.

[0018] In a further aspect the invention concerns novel compounds offormula (I) as defined above, with the proviso that5-methyl-3-phenyl-7-(phenylmethoxy)-pyrazolo[1,5-a]-pyrimidine and2,5-dimethyl-7-(methylamino)-3-phenyl-pyrazolo[1,5-a]pyrimidine are notincluded

[0019] The proviso is intended to exclude compounds disclosed inJP-61/057,587 and JP-42/011,753.

[0020] As used in the foregoing definitions and hereinafter, halo isgeneric to fluoro, chloro, bromo and iodo; C₁₋₆alkanediyl definesbivalent straight and branched chained saturated hydrocarbon radicalshaving from 1 to 6 carbon atoms, such as, for example, methylene,1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl, 1,5-pentanediyl,1,6-hexanediyl and the branched isomers thereof; C₁₋₂alkyl definesstraight saturated hydrocarbon radicals having from 1 to 2 carbon atomssuch as methyl and ethyl; C₂₋₄alkyl defines straight and branched chainsaturated hydrocarbon radicals having from 2 to 4 carbon atoms such asethyl, propyl, butyl, 1-methylethyl and the like; C₃₋₄alkyl definesstraight and branched chain saturated hydrocarbon radicals having from 3to 4 carbon atoms such as propyl, butyl, 1-methylethyl and the like;C₁₋₆alkyl includes C₁₋₂alkyl and C₃₋₄alkyl radicals as definedhereinbefore and the higher homologs thereof having from 5 to 6 carbonatoms such as, pentyl, the pentyl isomers, hexyl and the hexyl isomers;C₁₋₈alkyl includes C₁₋₆alkyl and the higher homologues thereof havingfrom 7 to 8 carbon atoms such as, for example, heptyl, octyl and thelike; C₃₋₆alkenyl defines straight and branched chain hydrocarbonradicals containing one double bond and having from 3 to 6 carbon atomssuch as, for example, 2-propenyl, 3-butenyl, 2-pentenyl, 3-pentenyl,3-methyl-2-butenyl, and the like; and where said C₃₋₆alkenyl is linkedto a nitrogen or oxygen, the carbon atom making the link preferably issaturated. C₃₋₆cycloalkyl comprises cyclopropyl, cyclobutyl, cyclopentyland cyclohexyl. HydroxyC₁₋₆alkyl refers to C₁₋₆alkyl substituted with ahydroxyl-group. Homopiperidinyl refers to a 7 membered saturated ringcontaining one nitrogen atom.

[0021] Depending on the nature of some of the substituents, thecompounds of formula (I) may contain one or more asymmetric centerswhich may be designated with the generally used R and S nomenclature.

[0022] The compounds of the present invention contain basic nitrogenatoms and, as such, can be present as the free base or in the form ofacid addition salts, both being part of this invention. Acid additionsalts may be prepared by methods well known in the art, and may beformed from organic and inorganic acids. Suitable organic acids includemaleic, fumaric, benzoic, ascorbic, succinic, methanesulfonic, acetic,oxalic, propionic, tartaric, salicylic, citric, gluconic, lactic,mandelic, cinnamic, aspartic, stearic, palmitic, glycolic, glutamic, andbenzenesulfonic acids. Suitable inorganic acids include hydrochloric,hydrobromic, sulfuric, phosphoric, and nitric acids.

[0023] Particular groups of compounds within the invention are thosecompounds of formula (I) wherein one or more of the followingrestrictions apply:

[0024] a) R¹ is NR⁴R⁵ wherein R⁴ is hydrogen, C₁₋₆alkyl,hydroxyC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl or C₃₋₆alkenyl; inparticular C₂₋₄alkyl, hydroxyC₁₋₂alkyl, C₃₋₄alkenyl orC₁₋₂alkylcarbonyloxyC₂₋₄alkyl; and R⁵ is C₁₋₈alkyl, C₃₋₆alkenyl,hydroxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl, phenylmethyl orC₃₋₆cycloalkylmethyl; in particular C₂₋₄alkyl, C₃₋₄alkenyl,hydroxyC₂₋₄alkyl or cyclopropylmethyl;

[0025] b) R¹ is OR⁵ wherein R⁵ is C₁₋₆alkyl; in particular C₂₋₄alkyl;

[0026] c) R² is C₁₋₆alkyl; in particular C₁₋₂alkyl;

[0027] d) R³ is hydrogen, C₁₋₆alkyl or C₁₋₆alkylthio; in particularhydrogen, C₁₋₂alkyl or C₁₋₂alkylthio;

[0028] e) Ar is a phenyl substituted with 1, 2 or 3 substituents eachindependently selected from C₁₋₆alkyl, C₁₋₆alkyloxy or halo; wherein thephenyl moiety is preferably substituted in the 3-, 4-, 6-, 2,4- or2,4,6-positions; or Ar is a pyridinyl substituted with 1, 2 or 3substituents each independently selected from halo, amino, nitro,trifluoromethyl, mono- or di(C₁₋₆alkyl)amino, piperidinyl or C₁₋₆alkyl;wherein the pyridinyl moiety preferably is connected via the 2- or3-position to the remainder of the molecule.

[0029] Another particular group of compounds are those compounds offormula (I) wherein R¹ is NR⁴R⁵ and R⁴ and R⁵ are taken together withthe nitrogen atom to which they are attached to form a pyrrolidinyl,piperidinyl, homopiperidinyl or morpholinyl group, optionallysubstituted with C₁₋₆alkyl or C₁₋₆alkyloxyC₁₋₆alkyl.

[0030] Preferred compounds are those compounds of formula (I) wherein R¹is NR⁴R⁵ and R⁴ is C₃₋₄alkyl or allyl, preferably propyl; R⁵ isC₂₋₄alkyl, allyl or cyclopropylmethyl, preferably propyl; R² is methyl;R³ is hydrogen, methyl or methylthio, preferably propyl; and Ar is aphenyl substituted with 1, 2 or 3 substituents each independentlyselected from halo, methyl or methoxy; and Ar in particular is pyridinylsubstituted with 1, 2 or 3 substituents each independently selected fromhalo, methyl or dimethylamino.

[0031] More preferably Ar is 3-pyridinyl substituted in the 4- and/or6-position with methyl or dimethylamino.

[0032] Most preferred are those compounds selected from

[0033]3-(2,4-dichlorophenyl)-5-methyl-7-(N-propyl-N-cyclopropanemethylamino)-pyrazolo[2,3-a]pyrimidine;

[0034]3-(2,4-dichlorophenyl)-5-methyl-7-(N-allyl-N-cyclopropanemethylamino)-pyrazolo[2,3-a]pyrimidine;

[0035]2-methylthio-3-(2,4-dichlorophenyl)-5-methyl-7-(N,N-diallylamino)-pyrazolo[2,3-a]pyrimidine;

[0036]2-methylthio-3-(2,4-dichlorophenyl)-5-methyl-7-(N-butyl-N-cyclopropanemethylamino)pyrazolo[2,3-a]pyrimidine;

[0037]2-methylthio-3-(2,4-dichlorophenyl)-5-methyl-7-(N-propyl-N-cyclopropanemethylamino)pyrazolo[2,3-a]pyrimidine;

[0038]2-methyl-3-(4-chlorophenyl)-5-methyl-7-(N,N-dipropylamino)-pyrazolo[2,3-a]pyrimidine;

[0039]3-[6-(dimethylamino)-3-pyridinyl]-2,5-dimethyl-N,N-dipropylpyrazolo[2,3-a]pyrimidin-7-amine;or

[0040]3-[6-(dimethylamino)-4-methyl-3-pyridinyl]-2,5-dimethyl-N,N-dipropylpyrazolo[2,3-a]pyrimidine-7-amine;or

[0041]3-(2,4-dimethoxyphenyl)-2,5-dimethyl-7-(N-propyl-N-methyloxyethylamino)-pyrazolo[2,3-a]pyrimidine;

[0042] the stereoisomeric forms and the pharmaceutically acceptable acidaddition salts thereof.

[0043] The compounds of the present invention can generally be preparedby alkylating a pyrazolopyrimidine of formula (II) with an intermediateof formula (III). In intermediate (II), W is an appropriate leavinggroup such as halo, e.g. chloro, bromo, or a sulfonyloxy group, e.g. amesyloxy or a tosyloxy group.

[0044] The above reaction is typically conducted in a suitable solvent,e.g. an aprotic solvent such as DMF or acetonitrile, an ether, e.g.tetrahydrofuran, preferably at an elevated temperature and, when theintermediates of formula (III) are volatile amines, in a sealed reactionvial.

[0045] Also, compounds of formula (I) wherein R¹ is OR⁵, said compoundsbeing represented by formula (I-a), may be prepared by O-alkylating anintermediate of formula (VI) with an intermediate of formula (VII),wherein W is as defined above.

[0046] Said reaction for preparing compounds of formula (I-a) can beperformed in a reaction-inert solvent such as, for example,N,N-dimethylformamide, and in the presence of a suitable base such as,for example, sodium hydride, preferably at a temperature ranging betweenroom temperature and reflux temperature.

[0047] The compounds of formula (I) wherein R¹ is NR⁴R⁵, represented byformula (I-c), can be prepared from either compounds of formula (VIII)or (IX) by suitable N-alkylation reactions as depicted herebelow,wherein W is as previously defined. These N-alkylations are conducted ina reaction-inert solvent such as, for example, an ether e.g.tetrahydofuran and preferably in the presence of a strong base, e.g.NaH.

[0048] In certain instances, this reaction can give rise to sideproducts wherein R² is alkylated by (R⁴ or R⁵)—W, in particular where R²is methyl and R⁴ or R⁵ is C₁₋₆alkyl.

[0049] As outlined below, compounds of formula (I) may be converted intoeach other following art-known transformation procedures.

[0050] The compounds of formula (I) wherein R³ is C₁₋₆alkylthio can beconverted into compounds of formula (I) wherein R³ is C₁₋₆alkylsulfonylor C₁₋₆alkylsulfoxy by an oxidation reaction, e.g. treatment with aperoxide such as 3-chloroperbenzoic acid in a reaction-inert solvent,e.g. dichloromethane. By controlling the amount of oxidant and otherreaction parameters, either compounds of formula (I) wherein R³ isC₁₋₆alkylsulfonyl or C₁₋₆alkylsulfoxy can be obtained, or a mixture ofboth, which subsequently can be separated by conventional methods, e.g.column chromatography.

[0051] The compounds of formula (I) may also be converted into eachother via art-known reactions or functional group transformations. Forinstance, compounds of formula (I) bearing a hydroxyC₁₋₆alkyl group maybe converted into compounds of formula (I) bearing aC₁₋₆alkylcarbonyloxyC₁₋₆alkyl group, e.g. by treatment with an acidanhydride in an reaction-inert solvent such as, e.g. dichloromethane,and optionally in the presence of a base such as, e.g. pyridine.

[0052] Compounds of formula (I) bearing a nitro group may be convertedto compounds of formula (I) bearing an amino group and subsequently tocompounds of formula (I) having a mono- or di(C₁₋₆alkyl)amino group.Also, the amino group may be converted using a diazotization reaction toa halo.

[0053] Further, the Ar group of compounds of formula (I) can behalogenated using a halogenating agent such as, e.g. chlorine orbromine, in a suitable solvent, e.g. acetic acid, and optionally thereaction may be performed at a temperature ranging between roomtemperature and the reflux temperature of the reaction mixture.

[0054] Intermediates of formula (II) can be prepared according to theprocedure as described in Robins et al., J. Heterocyclic Chem.22:601-634, 1985.

[0055] Aminopyrazole derivatives (IV) are reacted with a β-keto ester(V), preferably under reflux conditions and in a suitable reaction-inertsolvent such as an ether, e.g. THF, yielding hydroxypyrazolopyrimidines(VI) which are converted into intermediates of formula (II) byconverting the hydroxy group of intermediate (VI) into leaving group W,e.g. by treating (VI) with methanesulfonyloxy chloride or a halogenatingreagent such as, e.g. POCl₃.

[0056] Intermediates of formula (VIII) are prepared by treatingintermediates of formula (II) with ammonia.

[0057] In an embodiment, this invention also provides for intermediatesof formula (II′), wherein W′ represents hydroxy, halo, mesyloxy ortosyloxy; with the proviso that Ar is other than phenyl.

[0058] Said intermediates of formula (II′) may be prepared according toprocedures used to prepare intermediates of formula (II), therebythereby yielding compounds of formula (II′-a), defined as compounds offormula (II′) wherein W′ is hydroxy; and optionally converting compoundsof formula (II′-a) into compounds of formula (II′-b), defined ascompounds of formula (II′) wherein W′ is other than hydroxy.

[0059] Stereoisomers may be prepared by separation of the end productsof formula (I) following art-known procedures, e.g. by treatment with anoptically active acid and separating the thus-formed diastereoisomericsalts by selective crystallization or column chromatography. Or,stereoisomers may be prepared by using stereoisomeric starting materialsin any of the above reaction schemes or in the preparation ofintermediates described hereinafter.

[0060] The effectiveness of a compound as a CRF receptor antagonist maybe determined by various assay methods. Suitable CRF antagonists of thisinvention are capable of inhibiting the specific binding of CRF to itsreceptor and antagonizing activities associated with CRF. A compound ofstructure (I) may be assessed for activity as a CRF antagonist by one ormore generally accepted assays for this purpose, including (but notlimited to) the assays disclosed by DeSouza et al. (J. Neuroscience7:88, 1987) and Battaglia et al. (Synapse 1:572, 1987). As mentionedabove, suitable CRF antagonists include compounds which demonstrate CRFreceptor affinity. CRF receptor affinity may be determined by bindingstudies that measure the ability of a compound to inhibit the binding ofa radiolabeled CRF (e.g. [¹²⁵I]tyrosine CFR) to receptor (e.g.,receptors prepared from rat cerebral cortex membranes). The radioligandbinding assay described by DeSouza et al. (supra, 1987) provides anassay for determining a compound's affinity for the CRF receptor. Suchactivity is typically calculated from the IC₅₀ as the concentration of acompound necessary to displace 50% of the radiolabeled ligand from thereceptor, and is reported as a “K_(i)” value calculated by the followingequation: $K_{i} = \frac{I\quad C_{50}}{1 + {L/K_{D}}}$

[0061] where L=radioligand and K_(D)=affinity of radioligand forreceptor (Cheng and Prusoff, Biochem. Pharmacol. 22:3099, 1973).

[0062] In addition to inhibiting CRF receptor binding, a compound's CRFreceptor antagonist activity may be established by the ability of thecompound to antagonize an activity associated with CRF. For example, CRFis known to stimulate various biochemical processes, including adenylatecyclase activity. Therefore, compounds may be evaluated as CRFantagonists by their ability to antagonize CRF-stimulated adenylatecyclase activity by, for example, measuring cAMP levels. TheCRF-stimulated adenylate cyclase activity assay described by Battagliaet al. (supra, 1987) provides an assay for determining a compound'sability to antagonize CRF activity. Accordingly, CRF receptor antagonistactivity may be determined by assay techniques which generally includean initial binding assay (such as disclosed by DeSouza (supra, 1987))followed by a cAMP screening protocol (such as disclosed by Battaglia(supra, 1987)). With reference to CRF receptor binding affinities, CRFreceptor antagonists of this invention have a K_(i) of less than 10 μM.In a preferred embodiment of this invention, a CRF receptor antagonisthas a K_(i) of less than 1 μM, and more preferably less than 0.25 μM(i.e., 250 nM).

[0063] The CRF receptor antagonists of the present invention demonstrateactivity at the CRF receptor site, and may be used as therapeutic agentsfor the treatment of a wide range of disorders or illnesses includingendocrine, psychiatric, and neurologic disorders or illnesses. Morespecifically, the CRF receptor antagonists of the present invention maybe useful in treating physiological conditions or disorders arising fromthe hypersecretion of CRF. Because CRF is believed to be a pivotalneurotransmitter that activates and coordinates the endocrine,behavioral and automatic responses to stress, the CRF receptorantagonists of the present invention can be used to treatneuropsychiatric disorders. Neuropsychiatric disorders which may betreatable by the CRF receptor antagonists of this invention includeaffective disorders such as depression; anxiety-related disorders suchas generalized anxiety disorder, panic disorder, obsessive-compulsivedisorder, abnormal aggression, cardiovascular abnormalities such asunstable angina and reactive hypertension; and feeding disorders such asanorexia nervosa, bulimia, and irritable bowel syndrome. CRF antagonistsmay also be useful in treating stress-induced immune suppressionassociated with various diseases states, as well as stroke. Other usesof the CRF antagonists of this invention include treatment ofinflammatory conditions (such as rheumatoid arthritis, uveitis, asthma,inflammatory bowel disease and G.I. motility), Cushing's disease,infantile spasms, epilepsy and other seizures in both infants andadults, and various substance abuse and withdrawal (includingalcoholism).

[0064] In another embodiment of the invention, pharmaceuticalcompositions containing one or more CRF receptor antagonists aredisclosed. For the purposes of administration, the compounds of thepresent invention may be formulated as pharmaceutical compositions.Pharmaceutical compositions of the present invention comprise a CRFreceptor antagonist of the present invention (i.e., a compound ofstructure (I)) and a pharmaceutically acceptable carrier and/or diluent.The CRF receptor antagonist is present in the composition in an amountwhich is effective to treat a particular disorder, that is, in an amountsufficient to achieve CRF receptor antagonist activity, and preferablywith acceptable toxicity to the patient. Preferably, the pharmaceuticalcompositions of the present invention may include a CRF receptorantagonist in an amount from 0.1 mg to 250 mg per dosage depending uponthe route of administration, and more preferably from 1 mg to 60 mg.Appropriate concentrations and dosages can be readily determined by oneskilled in the art.

[0065] Pharmaceutically acceptable carrier and/or diluents are familiarto those skilled in the art. For compositions formulated as liquidsolutions, acceptable carriers and/or diluents include saline andsterile water, and may optionally include antioxidants, buffers,bacteriostats and other common additives. The compositions can also beformulated as pills, capsules, granules, or tablets which contain, inaddition to a CRF receptor antagonist, diluents, dispersing and surfaceactive agents, binders, and lubricants. One skilled in this art mayfurther formulate the CRF receptor antagonist in an appropriate manner,and in accordance with accepted practices, such as those disclosed inRemington's Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co.,Easton, USA, 1990.

[0066] In another embodiment, the present invention provides a methodfor treating a variety of disorders or illnesses, including endocrine,psychiatric and neurologic disorders or illnesses. Such methods includeadministering of a compound of the present invention to a warm-bloodedanimal in an amount sufficient to treat the disorder or illness. Suchmethods include systemic administration of a CRF receptor antagonist ofthis invention, preferably in the form of a pharmaceutical composition.As used herein, systemic administration includes oral and parenteralmethods of administration. For oral administration, suitablepharmaceutical compositions of CRF receptor antagonists include powders,granules, pills, tablets, and capsules as well as liquids, syrups,suspensions, and emulsions. These compositions may also includeflavorants, preservatives, suspending, thickening and emulsifyingagents, and other pharmaceutically acceptable additives. For parentaladministration, the compounds of the present invention can be preparedin aqueous injection solutions which may contain, in addition to the CRFreceptor antagonist, buffers, antioxidants, bacteriostats, and otheradditives commonly employed in such solutions.

[0067] As mentioned above, administration of a compound of the presentinvention can be used to treat a wide variety of disorders or illnesses.In particular, the compounds of the present invention may beadministered to a warm-blooded animal for the treatment of depression,anxiety disorder, panic disorder, obsessive-compulsive disorder,abnormal aggression, unstable angina, reactive hypertension, anorexianervosa, bulimia, irritable bowel syndrome, stress-induced immunesuppression, stroke, inflammation, Cushing's disease, infantile spasms,epilepsy, and substance abuse or withdrawal.

[0068] Hence, this invention provides the use of compounds of formula(I) for the manufacture of a medicine for treating physiologicalconditions or disorders arising from the hypersecretion ofcorticotropin-releasing factor (CRF); and in a further embodiment theuse of novel compounds of formula (I) as a medicine is provided.

[0069] The following examples are provided for purposes of illustration,not limition.

EXPERIMENTAL PART

[0070] Hereinafter “THF” means tetrahydrofuran and “DCM” meansdichloromethane.

[0071] A. Preparation of the Intermediates.

EXAMPLE A.1

[0072] a) 3-Amino-4-(2,4-dichlorophenyl)pyrazole and ethyl acetoacetate(2 equivalents) were dissolved in dioxane and heated under refluxovernight. The mixture was concentrated in vacuo and diluted with ethylacetate. An off-white solid formed after 2 days standing was collectedby vacuum filtration, yielding3-(2,4-dichlorophenyl)-5-methyl-7-hydroxypyrazolo[2,3-a]pyrimidine(intermediate 1).

[0073] b) Intermediate 1 (300 mg) was mixed with POCl₃ (1.5 ml) andheated to reflux for 1 hour. The resultant purple solution was carefullytransferred into ice-water. The product was extracted with ethylacetate, washed with saturated sodium bicarbonate and brine, dried overMgSO₄ and concentrated in vacuo to give3-(2,4-dichlorophenyl-5-methyl-7-chloropyrazolo[2,3-a]pyrimidine(intermediate 2) as a brown solid (260 mg, 82%).

EXAMPLE A.2

[0074] a) To a stirred solution of sodium hydride (60%, 25 mmol) in THF(10 ml) 6-(dimethylamino)-3-pyridineacetonitrile was added dropwise (10mmol) in THF (10 ml). The solution was allowed to stir for 10 minutesbefore ethyl acetate (30 mmol) was added slowly. The resultingsuspension was stirred at room temperature for another hour. Thereaction mixture was concentrated under vacuum and dissolved in ethylacetate/methanol (1:1) and filtered through silica. The filtrate wasconcentrated, yielding (intermediate 7).

[0075] b) A mixture of intermediate 7 and hydrazine hydrobromide (100mmol) was dissolved in ethanol/water (9:1, total 100 ml) and refluxedfor 1 hour. The reaction mixture was concentrated and the residue waspartitioned between ethyl acetate and sodium bicarbonate solution. Thecombined extracts were dried over sodium sulfate, filtered andconcentrated to dryness. The residue was dissolved in 1,4-dioxane (200ml) and refluxed for 16 hours in the presence of ethyl acetoacetate. Thereaction mixture was cooled and an off-white solid precipitated out.Diethyl ether was added to aid in crystallization and the precipitatewas filtered off and dried, yielding3-[2-(dimethylamino)-5-pyridinyl]-2,5-dimethyl-7-hydroxypyrazolo[2,3-a]pyrimidine(intermediate 8).

[0076] c) Intermediate 8 was dissolved in POCl₃ (2 ml) and refluxed for2 hours. The reaction mixture was cooled and poured onto ice. Thesolution was made basic (pH=9) by addition of solid sodium carbonate andextracted with diethyl ether. The combined organic layers were driedover sodium sulfate, filtered and concentrated, yielding3-[6-(dimethylamino)-3-pyridinyl]-2,5-dimethyl-7-chloro-pyrazolo[2,3-a]pyrimidine(intermediate 9).

[0077] Table 1 lists the intermediates that were prepared according toone of the above Examples. TABLE 1

Ex. Interm. No. No. R³ Ar 2 A.1 H 2,4-dichlorophenyl 3 A.1 CH₃S2,4-dichlorophenyl 4 A.1 CH₃ 2,4-dichlorophenyl 5 A.1 H 4-chlorophenyl 6A.1 H 2,6-dichlorophenyl 13 A.1 CH₃ 4-chlorophenyl 14 A.1 CH₃3-methoxyphenyl 15 A.1 CH₃ 4-methoxyphenyl 16 A.1 CH₃2,4-dimethoxyphenyl 17 A.1 CH₃CH₂ 3,4-dimethoxyphenyl 18 A.1 H2,4,6-trimethoxyphenyl 9 A.2 CH₃ 6-dimethylamino-3-pyridinyl 10 A.2 CH₃6-dimethylamino-4-methyl-3-pyridinyl 11 A.2 CH₃6-methyl-5-nitro-2-pyridinyl 12 A.2 CH₃ 5-chloro-2-pyridinyl 19 A.1 CH₃6-methyl-3-pyridinyl 20 A.1 CH₃ 3-methyl-5-nitro-2-pyridinyl

[0078] B. Preparation of the Final Products.

EXAMPLE B.1

[0079] A mixture of intermediate 2 (21 mg) andN-propyl-N-cyclopropanemethylamine (75 mg) was heated in a sealedreaction vial at 100° C. overnight. Chromatography on a preparative TLCplate with 1:5 ethyl acetate-hexanes gave3-(2,4-dichlorophenyl)-5-methyl-7-(N-propyl-N-cyclopropanemethylamino)-pyrazolo[2,3-a]pyrimidine(compound 17) (17.4 mg) and3-(2,4-dichlorophenyl)-5-methyl-7-(N-cyclopropanemethylamino)-pyrazolo[2,3-a]pyrimidine(compound 8) (1 mg).

[0080] In a similar way, starting from intermediate 2 and(S)-(−)-leucinol respectively (R)-(+)-leucinol,(S)-2[[3-(2,4-dichlorophenyl)-5-methyl-7-pyrazolo[2,3-a]pyrimidinyl]amino]-4-methyl-1-pentanol(compound 11) and its R-analog (compound 12) were prepared.

EXAMPLE B.2

[0081] A solution of intermediate 10 (8 g) and di-n-propylamine (13 g)in acetonitrile (50 ml) was heated at reflux for 3 hours. The mixturewas filtrated through a short silica gel plug with ethyl acetate and thefiltrate was concentrated in vacuo to provide a light yellow solid whichwas recrystallized from ether-hexanes, yielding 8.9 g (93%) of3-[6-(dimethylamino)-4-methyl-3-pyridinyl]-2,5-dimethyl-N,N-dipropylpyrazolo[2,3-a]-pyrimidin-7-amine(compound 53).

[0082] Compound 53 was also converted to its hydrochloric acid additionsalt by dissolving compound 53 (8.1 g) in a mixture of diethyl ether(150 ml) and DCM (50 ml) and treating said mixture with HCl in diethylether (1 M, 21.3 ml) dropwise with stirring. The resulting off-whitesolid was collected by filtration, yielding 8.7 g (98%) of3-[6-(dimethylamino)-4-methyl-3-pyridinyl]-2,5-dimethyl-N,N-dipropyl-pyrazolo[2,3-a]pyrimidin-7-aminemonohydrochloride.

EXAMPLE B.3

[0083] Intermediate 3 (15 mg) was dissolved in ethanol (0.5 ml) andstirred in the presence of sodium ethoxide (12 mg) for 1 hour.Chromatography on silica gel gave3-(2,4-dichlorophenyl)-7-(ethoxy)-5-methyl-2-methylthio-pyrazolo[2,3-a]pyrimidine(compound 50).

EXAMPLE B.4

[0084] A solution of compound 6 (14 mg) in THF (3 ml) was treated withsodium hydride (60 mg, excess) at room temperature for 5 minutes,followed by iodopropane (0.3 ml). The reaction was stirred at roomtemperature overnight and quenched with methanol (0.5 ml). The resultantmixture was loaded onto a preparative TLC plate and developed with 1:5ethyl acetate-hexanes to give3-(2,4-dichlorophenyl)-5-methyl-7-[N-(3-methoxypropyl)-N-propylamino]-pyrazolo[2,3-a]pyrimidine(compound 45) (5.7 mg) as a colorless oil,3-(2,4-dichlorophenyl)-5-butyl-7-[N-(3-methoxypropyl)-N-propylamino]-pyrazolo[2,3-a]pyrimidine(compound 46) (4 mg) and3-(2,4-dichlorophenyl)-5-butyl-7-[3-methoxy-propylamino]-pyrazolo[2,3-a]pyrimidine(compound 47) (3.5 mg).

EXAMPLE B.5

[0085] Compound 26 (8 mg) was dissolved in DCM (1 ml) and treated withacetic anhydride (0.1 ml) and pyridine (0.1 ml). The mixture was stirredat room temperature overnight and concentrated in vacuo. The residue wasdiluted with ethyl acetate and filtrated through a short silica gelcolumn. Concentration of the filtrate gave3-(2,4-dichloro-phenyl)-5-methyl-7-[N-(2-acetoxyethyl)-N-benzylamino]-pyrazolo[2,3-a]-pyrimidine(compound 27) (8.6 mg) as a colorless oil.

EXAMPLE B.6

[0086] A solution of compound 40 (20 mg) in DCM (3 ml) was treated with3-chloroperbenzoic acid (19 mg). The solution was stirred at roomtemperature for 1 hour. Chromatography on a silica gel plate with ethylacetate-hexanes (1:1) yielded compound 43 and compound 44.

EXAMPLE B.7

[0087] A mixture of compound 55, palladium on activated carbon (100 mg)in dry ethanol (100 ml) was put on the hydrogenation unit andhydrogenation was carried out at 2.7 10⁵ Pa (40 psi) for 2 hours. Thereaction mixture was filtered and concentrated. The residue wasdissolved in diethyl ether and concentrated, yielding 1.49 g of3-(5-amino-3-methyl-2-pyridinyl)-2,5-dimethyl-N,N-dipropyl-pyrazolo[2,3-a]pyrimidine-7-amine(compound 56).

EXAMPLE B.8

[0088] To a stirring solution of compound 56 (1.39 g) and aqueousformaldehyde (6.4 g) in ACN (20 ml) was added sodium cyanoborohydride(743 mg) at 0° C. Glacial acetic acid (1 ml) was added and the reactionmixture was stirred at room temperature for 2 hours. The reactionmixture was partitioned between ethyl acetate and saturated aqueoussodium bicarbonate. The organic layer was washed with brine, dried oversodium sulfate, filtered, concentrated to dryness and purified by flashcolumn chromatography on silica gel (CH₂Cl₂/CH₃OH/NH₄OH 150:10: 1). Thedesired fraction was isolated, yielding 1.30 g of3-[5-(dimethylamino)-3-methyl-2-pyridinyl]-2,5-dimethyl-N,N-dipropyl-pyrazolo[2,3-a]pyrimidine-7-amine(compound 57).

EXAMPLE B.9

[0089] To a solution of compound 56 (100 mg) in a mixture ofconcentrated HCl (1 ml) and water (1 ml), cooled in an ice bath, wasadded a solution of sodium nitrite (21 mg) in water (1 ml). The solutionwas added to a mixture of copper(I)chloride (281 mg) in concentrated HClwith stirring in an ice bath. A solid separated and the mixture washeated to 60° C. and a clear solution was obtained. The reaction mixturewas basified with NaOH, extracted with ethyl acetate, washed with brineand concentrated. Purification by preparative TLC yielded 16 mg ofcompound 98.

EXAMPLE B.10

[0090] Compound 56 (100 mg) was added to a solution of fluoroboric acid(48% wt in water, 2 ml), cooled in an ice bath, followed by addition ofa solution of sodium nitrite (20 mg) in water (1 ml). The temperaturewas kept under 10° C. The solid was collected by filtration, dried andpartitioned between an aqueous sodium bicarbonate solution and ethylacetate. The organic layer was washed with brine, concentrated and theresidue was purified by preparative TLC yielding 20 mg of compound 100.

[0091] Tables 2 to 5 list the compounds that were prepared according toone of the above Examples and tables 6 and 7 list the analytical datafor these compounds. TABLE 2

Co. Ex. No. No. R⁴ R⁵ Ar 1 B.1 hydrogen n-propyl 2,4-dichlorophenyl 2B.1 hydrogen 2-methylpropyl 2,4-dichlorophenyl 3 B.1 hydrogen1,1-dimethylethyl 2,4-dichlorophenyl 4 B.1 hydrogen 3-hydroxypropyl2,4-dichlorophenyl 5 B.1 hydrogen 3-pentyl 2,4-dichlorophenyl 6 B.1hydrogen CH₃O(CH₂)₃— 2,4-dichlorophenyl 7 B.1 hydrogen (CH₃)₂CHO(CH₂)₃—2,4-dichlorophenyl 8 B.1 hydrogen cyclopropylmethyl 2,4-dichlorophenyl 9B.1 hydrogen 3-methyl-2-butyl 2,4-dichlorophenyl 10 B.1 hydrogen4-methyl-2-pentyl 2,4-dichlorophenyl 11 B.1 hydrogen

2,4-dichlorophenyl 12 B.1 hydrogen

2,4-dichlorophenyl 13 B.1 hydrogen

2,4-dichlorophenyl 14 B.3 methyl cyclopropylmethyl 2,4-dichlorophenyl 15B.1 ethyl n-butyl 2,4-dichlorophenyl 16 B.2 n-propyl n-propyl2,4-dichlorophenyl 17 B.1 n-propyl cyclopropylmethyl 2,4-dichlorophenyl18 B.1 n-propyl HOCH₂CH₂— 2,4-dichlorophenyl 19 B.1 n-propyl CH₃O(CH₂)₂—2,4-dichlorophenyl 20 B.2 n-propyl n-propyl 4-chlorophenyl 21 B.1n-propyl cyclopropyl 2,6-dichlorophenyl 22 B.1 n-propyl CH₃O(CH₂)₂2,6-dichlorophenyl 23 B.4 n-butyl cyclopropylmethyl 2,4-dichlorophenyl24 B.1 2-methylpropyl cyclopropylmethyl 2,4-dichlorophenyl 25 B.13-methyl-2-butyl cyclopropylmethyl 2,4-dichlorophenyl 26 B.1 HOCH₂CH₂—phenylmethyl 2,4-dichlorophenyl 27 B.5 CH₃COO(CH₂)₂— phenylmethyl2,4-dichlorophenyl 28 B.5 CH₃COO(CH₂)₂— n-propyl 2,4-dichlorophenyl 29B.4 allyl cyclopropylmethyl 2,4-dichlorophenyl 45 B.4 n-propylCH₃O(CH₂)₃ 2,4-dichlorophenyl 58 B.1 CH₃O(CH₂)₂— CH₃O(CH₂)₂—2,4-dichlorophenyl 59 B.1 hydrogen CH₃OCH₂CH(CH₃)— 2,4-dichlorophenyl 60B.1 hydrogen 1-hydroxy-2-hexyl 2,4-dichlorophenyl 61 B.1 hydrogen1-hydroxy-2-pentyl 2,4-dichlorophenyl 62 B.2 n-propyl n-propyl6-dimethylamino-2,4-dimethyl-3-pyridinyl 63 B.1 ethyl n-butyl6-dimethylamino-2,4-dimethyl-3-pyridinyl 64 B.1 n-propylcyclopropylmethyl 6-dimethylamino-2,4-dimethyl-3-pyridinyl 65 B.1n-propyl cyclopropylmethyl 6-methyl-3-pyridinyl 66 B.1 n-butyl n-butyl6-methyl-3-pyridinyl 67 B.1 n-propyl cyclopropylmethyl2,4-dimethoxyphenyl 68 B.2 n-propyl n-propyl 2,4,6-trimethoxyphenyl 69B.1 n-propyl CH₃O(CH₂)₂— 2,4,6-trimethoxyphenyl 70 B.1 n-propylcyclopropylmethyl 2,4,6-trimethoxyphenyl 71 B.1 ethyl n-butyl2,4,6-trimethoxyphenyl 72 B.1 ethyl ethyl 2,4,6-trimethoxyphenyl

[0092] TABLE 3

Co. Ex. No. No. R³ R⁴ R⁵ Ar 30 B.1 CH₃S hydrogen n-propyl2,4-dichlorophenyl 31 B.1 CH₃S hydrogen 2-propyl 2,4-dichlorophenyl 32B.1 CH₃S hydrogen 3-heptyl 2,4-dichlorophenyl 34 B.1 CH₃S hydrogen2-methoxyphenylmethyl 2,4-dichlorophenyl 35 B.1 CH₃S methyl methyl2,4-dichlorophenyl 36 B.1 CH₃S ethyl ethyl 2,4-dichlorophenyl 37 B.2CH₃S n-propyl n-propyl 2,4-dichlorophenyl 38 B.1 CH₃S n-propylcyclopropylmethyl 2,4-dichlorophenyl 39 B.1 CH₃S 2-propyl 2-propyl2,4-dichlorophenyl 40 B.1 CH₃S n-butyl n-butyl 2,4-dichlorophenyl 41 B.1CH₃S n-butyl cyclopropylmethyl 2,4-dichlorophenyl 42 B.1 CH₃S allylallyl 2,4-dichlorophenyl 43 B.6 CH₃SO n-propyl cyclopropylmethyl2,4-dichlorophenyl 44 B.6 CH₃SO₂ n-propyl cyclopropylmethyl2,4-dichlorophenyl 73 B.2 CH₃CH₂ n-propyl n-propyl 3,4-dimethoxy-phenyl

[0093] TABLE 4

Co. Ex. No. No. R⁴ R⁵ Ar  48 B.2 n-propyl n-propyl 4-chlorophenyl  49B.1 n-propyl cyclopropylmethyl 2,4-dichlorophenyl  51 B.1 n-propylCH₃O(CH₂)₂— 2,4-dimethoxyphenyl  52 B.2 n-propyl n-propyl6-dimethylamino-3-pyridinyl  53 B.2 n-propyl n-propyl6-dimethylamino-4-methyl-3-pyridinyl  54 B.2 n-propyl n-propyl5-chloro-2-pyridinyl  55 B.2 n-propyl n-propyl3-methyl-5-nitro-2-pyridinyl  56 B.7 n-propyl n-propyl5-amino-3-methyl-2-pyridinyl  57 B.8 n-propyl n-propyl5-dimethylamino-3-methyl-2-pyridinyl  74 B.1 CH₃O(CH₂)₂— CH₃O(CH₂)₂—4-chlorophenyl  75 B.1 HOCH₂CH₂— phenylmethyl 4-chlorophenyl  76 B.1hydrogen 1-hydroxy-2-hexyl 4-chlorophenyl  77 B.1 hydrogen1-hydroxy-2-pentyl 4-chlorophenyl  78 B.1 hydrogen CH₃S(CH₂)₂—4-chlorophenyl  79 B.1 hydrogen

4-chlorophenyl  80 B.1 hydrogen 1-hydroxy-2-hexyl6-dimethylamimo-4-methyl-3-pyridinyl  81 B.1 ethyl n-butyl6-dimethylamino-4-methyl-3-pyridinyl  82 B.1 n-propyl cyclopropylmethyl6-dimethylamino-4-methyl-3-pyridinyl  83 B.1 n-propyl phenylmethyl6-dimethylamino-4-methyl-3-pyridinyl  84 B.1 allyl allyl6-dimethylamino-4-methyl-3-pyridinyl  85 B.1 n-butyl n-butyl6-dimethylamino-4-methyl-3-pyridinyl  86 B.1 hydrogen 3-pentyl6-dimethylamino-4-methyl-3-pyridinyl  87 B.1 hydrogen 2-propyl6-dimethylamino-4-methyl-3-pyridinyl  88 B.1 hydrogen 4-methyl-2-pentyl6-dimethylamino-4-methyl-3-pyridinyl  89 B.1 methyl n-butyl6-dimethylamino-4-methyl-3-pyridinyl  90 B.2 n-propyl n-propyl6-dimethylamino-2-methyl-3-pyridinyl  91 B.1 n-propyl cyclopropylmethyl6-dimethylamino-2-methyl-3-pyridinyl  92 B.1 ethyl n-butyl6-dimethylamino-2-methyl-3-pyridinyl  93 B.1 n-butyl n-butyl6-dimethylamino-2-methyl-3-pyridinyl  94 B.1 n-propyl n-propyl4,6-dimethyl-3-pyridinyl  95 B.1 n-propyl cyclopropylmethyl4,6-dimethyl-3-pyridinyl  96 B.2 n-propyl n-propyl6-dimethylamino-2,4-dimethyl-3-pyridinyl  97 B.1 n-propylcyclopropylmethyl 6-dimethylamino-2,4-dimethyl-3-pyridinyl  98 B.9n-propyl n-propyl 5-chloro-3-methyl-2-pyridinyl  99 B.9 n-propyln-propyl 5-iodo-3-methyl-2-pyridinyl 100  B.10 n-propyl n-propyl5-fluoro-3-methyl-2-pyridinyl 101 B.2 n-propyl n-propyl3-chloro-5-trifluoromethyl-2-pyridinyl 102 B.1 CH₃O(CH₂)₂— CH₃O(CH₂)₂—3-chloro-5-trifluoromethyl-2-pyridinyl 103 B.2 n-propyl n-propyl3,5-dichloro-2-pyridinyl 104 B.1 n-propyl CH₃O(CH₂)₂—3,5-dichloro-2-pyridinyl 105 B.1 n-propyl cyclopropylmethyl3,5-dichloro-2-pyridinyl 106 B.2 n-propyl n-propyl3-methyl-5-methoxy-2-pyridinyl 107 B.8 n-propyl n-propyl5-diethylamino-3-methyl-2-pyridinyl  33 B.8 n-propyl n-propyl6-diethylamino-4-methyl-3-pyridinyl 108 B.8 n-propyl n-propyl5-N-piperidinyl-3-methyl-2-pyridinyl 109 B.2 n-propyl n-propyl5-methyl-3-nitro-2-pyridinyl 110 B.2 n-propyl n-propyl3-amino-5-methyl-2-pyridinyl 111 B.9 n-propyl n-propyl3-chloro-5-methyl-2-pyridinyl 112 B.2 n-propyl n-propyl3-methylamino-5-methyl-2-pyridinyl 113 B.2 n-propyl n-propyl3-dimethylamino-5-methyl-2-pyridinyl 114 B.2 n-propyl n-propyl2-methyl-5-pyridinyl 115 B.1 n-propyl cyclopropylmethyl4-isopropylphenyl 116 B.2 n-propyl n-propyl 3,4-dimethoxyphenyl 117 B.1n-propyl CH₃O(CH₂)₂— 3,4-dimethoxyphenyl 118 B.1 n-propyl cyclopropyl2,4-dimethoxyphenyl 119 B.1 hydrogen CH₃O(CH₂)₂— 2,4-dimethoxyphenyl 120B.1 n-propyl HO(CH₂)₂— 2,4-dimethoxyphenyl 121 B.8 n-propyl n-propyl5-methylamino-3-methyl-2-pyridinyl 122 B.2 n-propyl n-propyl2,4-dimethoxyphenyl 123 B.1 ethyl n-butyl 2,4-dimethoxyphenyl 124 B.1n-propyl 2-hydroxypropyl 2,4-dimethoxyphenyl 125 B.2 n-propyl n-propyl4-methoxyphenyl 126 B.2 n-propyl n-propyl 3-methoxyphenyl 127 B.1n-propyl cyclopropylmethyl 3-methoxyphenyl Co. Ex. No. No. R⁴ and R⁵taken together Ar 128 B.1

4-chlorophenyl 129 B.1

4-chlorophenyl 130 B.1

6-dimethylamino-4-methyl-3-pyridinyl 131 B.1

6-dimethylamino-4-methyl-3-pyridinyl

[0094] TABLE 5

Co. Ex. No. No. R¹ R² R³ Ar 46 B.4

n-C₄H₉ H 2,4-dichlorophenyl 47 B.4

n-C₄H₉ H 2,4-dichlorophenyl 50 B.3 CH₃CH₂O— CH₃ CH₃—S—2,4-dichlorophenyl

[0095] TABLE 6 Analytical data Co. MS No. ¹H NMR data (CDCl₃) M + 1 1 δ0.45 (m, 2H), 0.78 (m, 2H), 1.20 (m, 1H), 2.84 (s, 3H), 3.40 (d, 2H),335 5.98 (s, 1H), 7.41 (dd, 1H), 7.52 (d, 1H), 7.76 (d, 1H), 8.32 (s,1H) 2 δ 0.88 (d, 3H), 1.05 (d, 3H), 1.74 (m, 1H), 2.52 (s, 3H), 3.20 (t,2H), 5.87 349 (s, 1H), 6.40 (t, 1H), 7.30 (dd, 1H), 7.47 (d, 1H), 7.99(d, 1H), 8.42 (s, 1H) 3 δ 1.57 (s, 9H), 2.54 (s, 3H), 6.05 (s, 1H), 6.48(s, 1H), 7.31 (d, 1H), 7.47 349 (s, 1H), 7.99 (d, 1H), 8.39 (s, 1H) 4 δ1.97 (m, 2H), 2.43 (s, 3H), 3.62 (m, 2H), 3.75 (t, 3H), 4,00 (brs, 1H),351 6.18 (s, 1H), 7.39 (dd, 1H), 7.52 (d, 1H), 8.19 (d, 1H), 8.43 (s,1H) 5 δ 1.03 (t, 6H), 1.60-1.85 (m, 4H), 2.56 (s, 3H), 3.49 (m, 1H),5.92 (s, 1H), 363 6.19 (d, 1H), 7.35 (dd, 1H), 7.50 (dd, 1H), 8.01 (d,1H), 8.44 (s, 1H) 6 δ 2.02 (m, 2H), 2.51 (s, 3H), 3.40 (s, 3H), 3.51 (m,4H), 5.91 (s, 1H), 6.71 365 (t, 1H), 7.31 (dd, 1H), 7.47 (d, 1H), 7.99(d, 1H), 8.42 (s, 1H) 7 δ 1.23 (d, 6H), 2.00 (m, 2H), 2.52 (s, 1H), 3.51(dd, 2H), 3.59 (m, 2H), 393 5.90 (s, 1H), 6.92 (t, 1H), 7.31 (dd, 1H),7.46 (d, 1H), 8.00 (d, 1H), 8.41 (s, 1H) 8 δ 0.35 (m, 2H), 0.68 (m, 2H),1.25 (m, 1H), 2.52 (s, 3H), 3.23 (dd, 2H), 347 5.88 (s, 1H), 6.40 (t,1H), 7.32 (d, 1H), 7.48 (s, 1H), 7.99 (s, 1H), 8.43 (s, 1H) 9 δ 1.03 (d,3H), 1.06 (d, 3H), 1.31 (d, 3H), 1.95 (m, 1H), 2.52 (s, 3H), 3.54 363(m, 1H), 5.88 (s, 1H), 6.23 (d, 1H), 7.32 (d, 1H), 7.47 (s, 1H), 7.98(d, 1H), 8.41 (s, 1H) 10 δ 0.95 (d, 3H), 0.97 (d, 3H), 1.40-1.80 (m,3H), 2.53 (s, 3H), 3.76 377 (m, 1H), 5.89 (s, 1H), 6.12 (d, 1H), 7.32(d, 1H), 7.47 (s, 1H), 7.97 (d, 1H), 8.40 (s, 1H) 11 δ 0.94 (d, 3H),0.99 (d, 3H), 1.53 (m, 1H), 1.76 (m, 1H), 2.50 (s, 3H), 393 2.93 (brs,1H), 3.48 (m, 1H), 3.76 (m, 1H), 5.97 (s, 1H), 6.22 (d, 1H), 7.35 (dd,1H), 7.51 (d, 1H), 7.95 (d, 1H), 8.36 (s, 12 δ 0.92 (d, 3H), 0.97 (d,3H), 1.52 (m, 2H), 1.72 (m, 1H), 2.47 (s, 3H), 393 3.02 (brs, 1H), 3.42(m, 1H), 3.72 (m, 2H), 5.94 (s, 1H), 6.20 (d, 1H), 7.33 (d, 1H), 7.49(s, 1H), 7.92 (d, 1H), 8.33 (s, 13 δ 0.99 (d, 6H), 1.98 (m, 1H), 2.44(s, 3H), 3.22 (brs, 1H), 3.46 (m, 2H), 379 3.73 (m, 1H), 5.88 (s, 1H),6.31 (d, 1H), 7.32 (d, 1H), 7.48 (s, 1H), 7.92 (d, 1H), 8.32 (s, 1H) 14δ 0.21 (m, 2H), 0.54 (m, 2H), 1.10 (m, 1H), 2.53 (s, 3H), 3.28 (s, 3H),379 3.91 (d, 2H), 5.99 (s, 1H), 7.32 (d, 1H), 7.48 (s, 1H), 7.97 (d,1H), 8.43 (s, 1H) 15 δ 0.96 (t, 3H), 1.31 (t, 3H), 1.38 (m, 2H), 1.71(m, 2H), 2.49 (s, 3H), 3.74 377 (t, 2H), 3.65 (q, 2H), 5.90 (s, 1H),7.30 (dd, 1H), 7.46 (d, 1H), 7.97 (d, 1H), 8.40 (s, 1H) 16 δ 0.96 (t,6H), 1.73 (m, 4H), 2.49 (s, 3H), 3.73 (t, 4H), 5.88 (s, 1H), 7.33 377(dd, 1H), 7.46 (d, 1H), 7.98 (d, 1H), 8.40 (s, 1H) 17 δ 0.26 (m, 2H),0.58 (m, 2H), 0.98 (t, 3H), 1.18 (m, 1H), 1.76 (m, 2H), 389 2.53 (s,3H), 3.76 (d, 2H), 3.81 (t, 2H), 6.02 (s, 1H), 7.34 (dd, 1H), 7.49 (dd,1H), 8.44 (s, 1H) 18 δ 1.04 (t, 3H), 1.77 (m, 2H), 2.53 (s, 3H), 3.47(t, 2H), 3.98 (m, 2H), 4.06 379 (m, 2H), 5.52 (brs, 1H), 6.03 (s, 1H),7.33 (d, 1H), 7.48 (s, 1H), 7.93 (d, 1H), 8.39 (s, 1H) 19 δ 0.98 (t,3H), 1.76 (m, 2H), 2.50 (s, 3H), 3.35 (s, 3H), 3.67 (t, 2H), 3.72 393(m, 2H), 4.14 (m, 2H), 5.95 (s, 3H), 7.29 (d, 1H), 7.47 (s, 1H), 7.97(d, 1H), 8.39 (s, 1H) 20 δ 0.96 (t, 6H), 1.73 (m, 4H), 2.55 (s, 3H),3.74 (t, 4H), 5.90 (s, 1H), 7.40 343 (d, 2H), 8.04 (d, 2H), 8.25 (s, 1H)21 δ 0.26 (m, 2H), 0.51 (m, 2H), 0.92 (t, 3H), 1.10 (m, 1H), 1.69 (m,2H), 389 2.72 (s, 3H), 3.41 (d, 2H), 3.47 (t, 2H), 6.17 (s, 1H), 7.18(t, 1H), 7.43 (d, 2H), 7.99 (s, 1H) 22 δ 0.93 (t, 3H), 1.67 (m, 2H),2.71 (s, 3H), 3.32 (s, 3H), 3.44 (t, 2H), 3.61 393 (m, 2H), 3.68 (m,2H), 6.16 (s, 1H), 7.17 (t, 1H), 7.41 (d, 2H), 7.99 (s, 1H) 23 δ 0.24(m, 2H), 0.55 (m, 2H), 0.95 (t, 3H), 1.38 (m, 2H), 1.69 (m, 2H), 4032.51 (s, 3H), 3.74 (d, 2H), 3.82 (t, 2H), 6.00 (s, 1H), 7.31 (dd, 1H),7.48 (dd, 1H), 7.98 (d, 1H), 8.42 (s, 1H) 24 δ 0.28 (m, 2H), 0.50 (m,2H), 0.94 (d, 6H), 1.10 (m, 1H), 2.05 (m, 1H), 403 2.52 (s, 3H), 3.62(d, 2H), 3.82 (d, 2H), 6.03 (s, 1H), 7.32 (d, 1H), 7.47 (s, 1H), 7.99(d, 1H), 8.43 (s, 1H) 25 δ 0.17 (m, 2H), 0.47 (m, 2H), 0.97 (t, 6H),1.05 (m, 1H), 1.67 (m, 4H), 417 2.52 (s, 3H), 2.43 (d, 2H), 4.71 (m,1H), 6.13 (s, 1H), 7.32 (dd, 1H), 7.47 (d, 1H), 8.02 (d, 1H), 8.43 (s,1H) 26 δ 2.41 (s, 3H), 4.01 (m, 2H), 4.15 (m, 2H), 4.79 (s, 2H), 5.62(brs, 1H), 427 6.02 (s, 1H), 7.35 (m, 6H), 7.48 (s, 1H), 7.91 (d, 1H),8.41 (s, 1H) 27 δ 1.91 (s, 3H), 2.47 (s, 3H), 4.20 (t, 2H), 4.43 (t,2H), 4.96 (s, 2H), 6.03 469 (s, 1H), 7.34 (m, 6H), 7.49 (s, 1H), 7.94(d, 1H), 8.44 (s, 1H) 28 δ 0.98 (t, 3H), 1.74 (m, 2H), 1.93 (s, 3H),2.49 (s, 3H), 3.59 (m, 2H), 4.84 421 (m, 2H), 4.39 (m, 2H), 5.95 (s,1H), 7.30 (dd, 1H), 7.46 (d, 1H), 7.92 (d, 1H), 8.38 (s, 1H) 29 δ 0.28(m, 2H), 0.60 (m, 2H), 1.20 (m, 1H), 2.51 (s, 3H), 3.70 (d, 2H), 3874.50 (d, 2H), 5.29 (d, 1H), 5.30 (d, 1H), 5.96 (m, 1H), 6.05 (s, 1H),7.32 (dd, 1H), 7.48 (d, 1H), 7.96 (d,1H), 8.44 (s, 1H) 30 δ 1.07 (t,3H), 1.80 (m, 2H), 2.46 (s, 3H), 2.55 (s, 3H), 3.36 (dt, 2H), 5.82 381(s, 1H), 6.29 (t, 1H), 7.29 (d, 1H), 7.38 (d, 1H), 7.51 (s, 1H) 31 δ1.41 (d, 6H), 2.46 (s, 3H), 2.55 (s, 3H), 3.85 (m, 1H), 5.82 (s, 1H),6.08 381 (d, 1H), 7.29 (d, 1H), 7.39 (d, 1H), 7.51 (s, 1H) 32 δ 0.92 (t,3H), 1.01 (t, 3H), 1.25-1.80 (m, 8H), 2.45 (s, 3H), 2.54 (s, 3H), 4373.48 (m, 1H), 5.79 (s, 1H), 6.02 (d, 1H), 7.27 (d, 1H), 7.39 (d, 1H),7.50 (s, 1H) 33 δ 0.96 (t, 3H), 1.20 (t, 6H), 1.74 (m, 4H), 2.15 (s,3H), 2.34 (s, 3H), 2.41 — (s, 3H), 3.56 (t, 4H), 3.79 (t, 4H), 5.77 (s,1H), 6.41 (s, 1H), 7.54 (s, 1H) 34 δ 2.45 (s, 3H), 2.56 (s, 3H), 3.93(s, 3H), 4.60 (d, 2H), 5.90 (s, 1H), 6.70 459 (t, 1H), 6.99 (t, 2H),7.27-7.41 (m, 4H), 7.52 (s, 1H) 35 δ 2.44 (s, 3H), 2.60 (s, 3H), 3.35(s, 6H), 5.00 (brs, 1H), 5.84 (s, 1H), 7.29 367 (d, 1H), 7.38 (d, 1H),7.51 (s, 1H) 36 δ 1.36 (s, 6H), 2.42 (s, 3H), 2.59 (s, 3H), 3.80 (m,4H), 5.80 (s, 1H), 7.29 395 (d, 1H), 7.38 (d, 1H), 7.51 (s, 1H) 37 δ0.98 (t, 6H), 1.78 (m, 4H), 2.40 (s, 3H), 2.59 (s, 3H), 3.69 (m, 4H),5.74 423 (s, 1H), 7.28 (d, 1H), 7.48 (d, 1H) 38 δ 0.30 (m, 2H), 0.62 (m,2H), 0.99 (t, 3H), 1.20 (m, 1H), 1.81 (m, 2H), 435 2.49 (s, 3H), 2.59(s, 3H), 3.65-3.90 (m, 4H), 5.87 (s, 1H), 7.29 (d, 1H), 7.39 (d, 1H),7.51 (s, 1H) 39 δ 1.05 (d, 3H), 1.38 (d, 3H), 2.45 (s, 3H), 2.56 (s,3H), 3.40 (m, 1H), 5.80 423 (s, 1H), 7.30 (d, 1H), 7.37 (d, 1H), 7.51(s, 1H) 40 δ 0.98 (t, 6H), 1.40 (m, 4H), 1.75 (m, 4H), 2.42 (s, 3H),2.59 (s, 3H), 3.71 451 (m, 4H), 5.76 (s, 1H), 7.29 (d, 1H), 7.39 (d,1H), 7.51 (s, 1H) 41 δ 0.29 (m, 2H), 0.61 (m, 2H), 0.93 (t, 3H),1.20-1.56 (m, 3H), 1.75 449 (m, 2H), 2.43 (s, 3H), 2.59 (s, 3H),3.65-3.90 (m, 4H), 5.88 (s, 1H), 7.30 (d, 1H), 7.39 (d, 1H), 7.51 (s,1H) 42 δ 2.42 (s, 3H), 2.59 (s, 3H), 4.34 (d, 4H), 5.30 (m, 4H), 5.87(s, 1H), 6.05 419 (m, 2H), 7.29 (d, 1H), 7.39 (d, 1H), 7.51 (s, 1H) 43 δ0.31 (m, 2H), 0.62 (m, 2H), 1.00 (t, 3H), 1.23 (m, 1H), 1.81 (m, 1H),451 2.49 (s, 3H), 3.05 (brs, 3H), 3.42-3.95 (m, 4H), 6.04 (s, 1H), 7.33(d, 1H), 7.50 (s, 1H), 7.51 (d, 1H) 44 δ 0.13 (m, 2H), 0.48 (m, 2H),0.90 (t, 3H), 1.05 (m, 1H), 1.60 (m, 2H), 485 (+H₂O) 2.64 (s, 3H), 3.05(brs, 3H), 3.48 (d, 2H), 3.71 (m, 2H), 6.04 (s, 1H), 7.37 (d, 1H), 7.54(s, 1H), 7.60 (d, 1H) 45 δ 0.95 (t, 3H), 1.42 (m, 2H), 1.98 (m, 2H),2.49 (s, 3H), 3.33 (s, 3H), 3.56 407 (m, 2H), 3.75 (m, 2H), 5.93 (m,1H), 7.31 (d, 1H), 7.46 (s, 1H), 7.96 (d, 1H), 8.40 (s, 1H) 46 δ 0.91(t, 3H), 0.95 (t, 3H), 1.35-2.05 (m, 8H), 2.70 (t, 2H), 3.33 (s, 3H),449 3.40 (m, 6H), 5.92 (s, 1H), 7.30 (d, 1H), 7.46 (s, 1H), 8.02 (d,1H), 8.41 (s, 1H) 47 δ 0.87 (t, 3H), 1.25 (m, 2H), 1.70 (m, 2H), 2.04(m, 2H), 2.68 (m, 2H), 407 3.41 (s, 3H), 3.56 (m, 4H), 5.86 (s, 1H),6.60 (t, 1H), 7.31 (d, 1H), 7.46 (s, 1H), 8.14 (d, 1H), 8.50 (s, 1H) 48δ 0.95 (t, 6H), 1.71 (m, 4H), 2.47 (s, 3H), 2.55 (s, 3H), 3.71 (t, 4H),5.82 357 (s, 1H), 7.39 (d, 2H), 7.70 (d, 2H) 49 δ 0.24 (m, 2H), 0.54 (m,2H), 0.96 (t, 3H), 1.12 (m, 1H), 1.74 (m, 2H), 403 2.35 (s, 3H), 2.44(s, 3H), 3.73 (d, 2H), 3.76 (t, 2H), 5.93 (s, 1H), 7.29 (d, 1H), 7.35(d, 1H), 7.51 (s, 1H) 50 δ 1.64 (t, 3H), 2.52 (s, 3H), 2.60 (s, 3H),4.56 (q, 2H), 6.02 (s, 3H), 7.30 368 (d, 1H), 7.37 (d, 1H), 7.51 (s, 1H)52 δ 0.9 (t, 6H), 1.68 (m, 4H), 2.13 (s, 3H), 2.30 (s, 3H), 2.37 (s,3H), M⁺ = 366 3.05 (s, 6H), 3.65 (m, 4H), 5.74 (s, 1H), 6.44 (s, 1H),8.01 (s, 1H) 53 δ 0.9 (t, 6H), 1.70 (m, 4H), 2.35 (s, 3H), 2.45 (s, 3H),3.05 (s, 6H), M⁺ = 380 3.65 (m, 4H), 5.74 (s, 1H), 6.44 (s, 1H), 8.01(s, 1H) 54 δ 0.95 (t, 6H), 1.73 (m, 4H), 2.52 (s, 3H), 2.76 (s, 3H),3.73 (t, 4H), — 5.85 (s, 1H), 7.68 (dd, 1H), 8.45 (d, 1H), 8.57 (d, 1H)55 δ 0.96 (t, 6H), 1.70-1.78 (m, 4H), 2.44 (s, 3H), 2.47 (s, 3H), 2.50(s, 383 3H), 3.73 (t, 4H), 5.48 (s, 1H), 8.36 (d, 1H), 9.32 (d, 1H) 56 δ0.93 (t, 6H), 1.66-1.73 (m, 4H), 2.19 (s, 3H), 2.36 (s, 3H), 2.42 (s,353 3H), 3.71 (t, 4H), 5.79 (s, 1H), 6.39 (d, 1H), 8.05 (d, 1H) 57 δ0.96 (t, 6H), 1.62-1.70 (m, 4H), 2.2 (s, 3H), 2.36 (s, 3H), 2.40 (s, 3813H), 2.96 (s, 6H), 3.68 (t, 4H), 5.76 (s, 1H), 6.91 (d, 1H), 8.08 (d,1H)

[0096] TABLE 7 Analytical data Co. No. Mass spectral data 58 409 [M⁺] 59365 [M⁺] 60 394 [MH⁺] 61 380 [MH⁺] 62 381 [MH⁺] 63 381 [MH⁺] 64 393[MH⁺] 65 350 [MH⁺] 66 366 [MH⁺] 67 381 [MH⁺] 68 399 [MH⁺] 69 415 [MH⁺]70 411 [MH⁺] 71 399 [MH⁺] 72 371 [MH⁺] 73 397 [MH⁺] 74 388 [M⁺] 75 407[MH⁺] 76 373 [MH⁺] 77 359 [MH⁺] 78 346 [M⁺] 79 373 [MH⁺] 80 397 [MH⁺] 81381 [MH⁺] 82 393 [MH⁺] 83 429 [MH⁺] 84 377 [MH⁺] 85 409 [MH⁺] 86 367[MH⁺] 87 339 [MH⁺] 88 381 [MH⁺] 89 367 [MH⁺] 90 381 [MH⁺] 91 393 [MH⁺]92 381 [MH⁺] 93 409 [MH⁺] 94 352 [MH⁺] 95 364 [MH⁺] 96 395 [MH⁺] 97 407[MH⁺] 98 372 [MH⁺] (Cl³⁵) 99 464 [MH⁺] 100 356 [MH⁺] 101 426 [MH⁺](Cl³⁵) 102 458 [MH⁺] (Cl³⁵) 103 392 [MH⁺] (Cl³⁵) 104 408 [MH⁺] (Cl³⁵)105 404 [MH⁺] (Cl³⁵) 106 368 [MH⁺] 107 409 [MH⁺] 108 421 [MH⁺] 109 383[MH⁺] 110 353 [MH⁺] 111 372 [MH⁺] (Cl³⁵) 112 366 [MH⁺] 113 381 [MH⁺] 114338 [MH⁺] 115 377 [MH⁺] 116 383 [MH⁺] 117 399 [MH⁺] 118 395 [MH⁺] 119415 [MH⁺] 120 385 [MH⁺] 121 367 [MH⁺] 122 383 [MH⁺] 123 383 [MH⁺] 124399 [MH⁺] 125 338 [MH⁺] 126 353 [MH⁺] 127 365 [MH⁺] 128 370 [M⁺] 129 368[M⁺] 130 393 [MH⁺] 131 379 [MH⁺]

[0097] C. Pharmacological Examples

EXAMPLE C.1 Representative Compounds Having CRF Receptor BindingActivity

[0098] Compounds were evaluated for binding activity to the CRF receptorby a standard radioligand binding assay as generally described byDeSouza et al. (J. Neurosci. 7:88-100, 1987). By utilizing variousradiolabeled CRF ligands, the assay may be used to evaluate the bindingactivity of the compounds of the present invention with any CRF receptorsubtype. Briefly, the binding assay involves the displacement of aradiolabeled CRF ligand from the CRF receptor.

[0099] More specifically, the binding assay was performed in 1.5 mlEppendorf tubes using approximately 1×10⁶ cells per tube stablytransfected with human CRF receptors. Each tube received about 0.1 ml ofassay buffer (e.g., Dulbecco's phosphate buffered saline, 10 mMmagnesium chloride, 20 μM bacitracin) with or without unlabeledsauvagine, urotensin I or CRF (final concentration, 1 μM) to determinenonspecific binding, 0.1 ml of [¹²⁵I] tyrosine—ovine CRF (finalconcentration ˜200 pM or approximately the K_(D) as determined byScatchard analysis) and 0.1 ml of a membrane suspension of cellscontaining the CRF receptor. The mixture was incubated for 2 hours at22° C. followed by the separation of the bound and free radioligand bycentrifugation. Following two washes of the pellets, the tubes were cutjust above the pellet and monitored in a gamma counter for radioactivityat approximately 80% efficiency. All radioligand binding data wasanalyzed using a non-linear least-square curve-fitting program.

[0100] Binding activity corresponds to the concentration (nM) of thecompound necessary to displace 50% of the radiolabeled ligand from thereceptor. The following compounds have K_(i)≦250 nM :5, 10, 12, 15-17,19, 23, 24, 26, 27, 29, 31-33, 36-43, 48, 49, 51-54 and 57-130 as listedin Tables 2-5. Compounds 17, 29, 38, 41, 42, 48, 49, 51-54, 57, 58, 70,71, 81-85, 90-92, 101, 103-105, 115, 118, 121-123 and 125 were found toshow the best score in this test.

EXAMPLE C.2 CRF Stimulated Adenylate Cyclase Activity

[0101] The compounds of the present invention may also be evaluated byvarious functional testing. For example, the compounds of the presentinvention may be screened for CRF-stimulated adenylate cyclase activity.An assay for the determination of CRF-stimulated adenylate cyclaseactivity may be performed as generally described by Battaglia et al.(Synapse 1:572, 1987), with modifications to adapt the assay to wholecell preparations.

[0102] More specifically, the standard assay mixture may contain thefollowing in a final volume of 0.5 ml: 2 mM L-glutamine, 20 mM HEPES,and 1 mM IMBX in DMEM buffer. In stimulation studies, whole cells withthe transfected CRF receptors are plated in 24-well plates and incubatedfor 1 hour at 37° C. with various concentrations of CRF-related andunrelated peptides in order to establish the pharmacological rank-orderprofile of the particular receptor subtype. Following the incubation,the medium is aspirated, the wells rinsed once gently with fresh medium,and the medium aspirated. To determine the amount of intracellular cAMP,300 μl of a solution of 95% ethanol and 20 mM aqueous hydrochloric acidis added to each well and the resulting suspensions are incubated at−20° C. for 16 to 18 hours. The solution is removed into 1.5 mlEppendorf tubes and the wells washed with an additional 200 μl ofethanol/aqueous hydrochloric acid and pooled with the first fraction.The samples are lyophilized and then resuspended with 500 μl sodiumacetate buffer. The measurement of cAMP in the samples is performedusing a single antibody kit. For the functional assessment of thecompounds, a single concentration of CRF or related peptides causing 80%stimulation of cAMP production is incubated along with variousconcentrations of competing compounds (10⁻¹² to 10⁻⁶M).

EXAMPLE C.3

[0103] The plus-maze and defensive withdrawal paradigms are correlatedmeasures of exploratory activity sensitive to anxiogenic and anxiolyticeffects of experimental treatments. These animal models are employed toexamine anxiolytic and anti-stress actions of compounds of the presentinvention.

EXAMPLE C.3-a

[0104] The Elevated Plus-maze Paradigm

[0105] This test predicts how animals respond to an approach-avoidancesituation involving a bright lighted space versus a dark “safe” area.Both spaces are elevated off the ground and constitute two runwaysintersecting in the form of plus sign. This type of approach-avoidancesituation is a classical test of “emotionality” and reactivity and isvery sensitive to treatments that produce disinhibition (such ansedative/hypnotic drugs) and stress. No motivational constraints arenecessary and the animal is free to remain in the dark or venture out onthe open arms. The plus-maze apparatus has four arms (10×50 cm) at rightangles to each other and is elevated from the floor (50 cm). Two of thearms are enclosed with walls (40 cm high) and two arms have no walls(open arms). Subjects are placed individually onto the center of themaze and allowed free access to all four arms for 5 minutes. Subjectsare observed through a window in the door and via an on-line display ofthe rat's location on a computer monitor. Time spent in each arm isrecorded automatically by photocell beams and a computer program. Themaze is wiped clean with a damp cloth between each trial. The measure ofanxiogenic-like behavior in this task is a decrease in time spent onopen arms while the measure of anti-stress efficacy is a complementaryincrease in time spent on open arms.

[0106] Validation of the Plus-maze Using CRF Peptides:

[0107] Central administration of CRF and exposure to any of severalexperimental stressors suppresses exploration in the elevated plus mazemodel of anxiety. When measuring the behavioral response to socialdefeat, central administration of the alpha-helical CRF (9-41)antagonist either post-stress [25 μg ICV] or pre-stress [1 μg ICV]reverses the anxiogenic effect of social defeat. This anti-stress actionof the CFR antagonist is also exerted following intracerebraladministration into the central nucleus of amygdala [250 ng IC].

[0108] Rats were administered the test compounds orally one hour priorto the five minute test. Some groups were placed in a water-filled poolfor ninety seconds immediately prior to placement on the Plus-Maze(Stress group) while control subjects were removed directly from thehome cage (Non-Stress group). In the non-treated animal group asignificant reduction of the percentage of the average time spent in theopen arms was observed (from about 36 to about 16%). Upon administrationof 2.5 or 20 mg/kg of compound 53, the % time in the open arms returnedto a level equal, within the error range, to the untreated group.

EXAMPLE C.3-b

[0109] The Defensive Withdrawal Paradigm

[0110] Testing takes place in a plexiglas open field (106×92×77 cm)containing a cylindrical galvanized steel chamber measuring 17.1 cm deepand 10.2 cm in diameter. The chamber is open at one end and is locatedalong the wall of the open field aligned lengthwise and 15.0 cm awayfrom a corner of the open field. The open end faces the corner. The openfield is illuminated by fluorescent ceiling lighting. For testing, theanimals are introduced into the unfamiliar test environment by placingthem into the small chamber. Tests take 5 minutes in duration and theapparatus is cleaned with a mild acetic acid solution after each test.The test compound is administered orally one hour before the 5 minutestest. The behavior of the animals is monitored and recorded by videocamera. The latency to leave the chamber will be measured and defined asthe placement of all four paws in the open field. Also measured is thenumber of passages made between the chamber and the open field and theaverage length of time in the chamber per entry. The measure ofanxiolytic efficacy is a decrease in mean time spent within the enclosedchamber. Compound 53 reduced the mean time in the chamber from about 80seconds to about 20 to 40 seconds when administered at doses of 0.63,2.5 and 20 mg/kg p.o. to rats.

[0111] Validation of Defensive Withdrawal Using CRF Peptides

[0112] When injected ICV, both alpha-helical CFR (9-41) and CRF modifybehavior in the defensive withdrawal paradigm. In particular, ICVadministration of CRF in animals previously familiarized with theapparatus increases both the latency to emerge from the small chamberand mean time spent in the chamber over the fifteen minute session.Similarly, infusion of CRF into the locus ceruleus produced similarchanges in defensive withdrawal behavior suggesting that the interactionof CRF with noradrenergic neurons is involved in defensive withdrawalbehavior in rats. Conversely, ICV administration of alpha-helical CRF(9-14) or d-Phe CRF (12-41), competitive CRF receptor antagonists,reverses the CRF-like effect of a restraint stressor in familiarenvironment condition and significantly decreases latency to emerge andmean time in chamber measures in animals unfamiliar with the apparatus.

[0113] D. Composition Examples

[0114] The following formulations exemplify typical pharmaceuticalcompositions in dosage unit form suitable for systemic or topicaladministration to warm-blooded animals in accordance with the presentinvention.

[0115] “Active ingredient” (A.I.) as used throughout these examplesrelates to a compound of formula (I), a N-oxide form, a pharmaceuticallyacceptable acid or base addition salt or a stereochemically isomericform thereof.

EXAMPLE D.1

[0116] Oral Solutions

[0117] 9 g of methyl 4-hydroxybenzoate and 1 g of propyl4-hydroxybenzoate are dissolved in 41 of boiling purified water. In 31of this solution are dissolved first 10 g of 2,3-dihydroxybutanedioicacid and thereafter 20 g of the A.I. The latter solution is combinedwith the remaining part of the former solution and 121 of1,2,3-propanetriol and 31 of sorbitol 70% solution are added thereto. 40g of sodium saccharin are dissolved in 0.5 l of water and 2 ml ofraspberry and 2 ml of gooseberry essence are added. The latter solutionis combined with the former, water is added q.s. to a volume of 201providing an oral solution comprising 5 mg of the A.I. per teaspoonful(5 ml). The resulting solution is filled in suitable containers.

EXAMPLE D.2

[0118] Capsules

[0119] 20 g of the A.I., 6 g sodium lauryl sulfate, 56 g starch, 56 glactose, 0.8 g colloidal silicon dioxide, and 1.2 g magnesium stearateare vigorously stirred together. The resulting mixture is subsequentlyfilled into 1000 suitable hardened gelatin capsules, each comprising 20mg of the A.I.

EXAMPLE D.3

[0120] Film-coated Tablets

[0121] Preparation of Tablet Core

[0122] A mixture of 100 g of the A.I., 570 g lactose and 200 g starch ismixed well and thereafter humidified with a solution of 5 g sodiumdodecyl sulfate and 10 g polyvinyl-pyrrolidone in about 200 ml of water.The wet powder mixture is sieved, dried and sieved again. Then there areadded 100 g microcrystalline cellulose and 15 g hydrogenated vegetableoil. The whole is mixed well and compressed into tablets, giving 10.000tablets, each comprising 10 mg of the active ingredient.

[0123] Coating

[0124] To a solution of 10 g methyl cellulose in 75 ml of denaturatedethanol there is added a solution of 5 g of ethyl cellulose in 150 ml ofdichloromethane. Then there are added 75 ml of dichloromethane and 2.5ml 1,2,3-propanetriol. 10 g of polyethylene glycol is molten anddissolved in 75 ml of dichloromethane. The latter solution is added tothe former and then there are added 2.5 g of magnesium octadecanoate, 5g of polyvinylpyrrolidone and 30 ml of concentrated colour suspensionand the whole is homogenated. The tablet cores are coated with the thusobtained mixture in a coating apparatus.

EXAMPLE D.4

[0125] Injectable Solution

[0126] 1.8 g methyl 4-hydroxybenzoate and 0.2 g propyl 4-hydroxybenzoatewere dissolved in about 0.51 of boiling water for injection. Aftercooling to about 50° C. there were added while stirring 4 g lactic acid,0.05 g propylene glycol and 4 g of the A.I. The solution was cooled toroom temperature and supplemented with water for injection q.s. ad 11volume, giving a solution of 4 mg/ml of A.I. The solution was sterilizedby filtration and filled in sterile containers.

EXAMPLE D.5

[0127] Suppositories

[0128] 3 Grams A.I. was dissolved in a solution of 3 grams2,3-dihydroxybutanedioic acid in 25 ml polyethylene glycol 400.12 Gramssurfactant and 300 grams triglycerides were molten together. The lattermixture was mixed well with the former solution. The thus obtainedmixture was poured into moulds at a temperature of 37 to 38° C. to form100 suppositories each containing 30 mg/ml of the A.I.

1. A compound of formula

including the stereoisomers and the pharmaceutically acceptable acidaddition salt forms thereof, wherein R¹ is NR⁴R⁵ or OR⁵; R² isC₁₋₆alkyl, C₁₋₆alkyloxy or C₁₋₆alkylthio; R³ is hydrogen, C₁₋₆alkyl,C₁₋₆alkylsulfonyl, C₁₋₆alkylsulfoxy or C₁₋₆alkylthio; R⁴ is hydrogen,C₁₋₆alkyl, mono- or di(C₃₋₆cycloalkyl)methyl, C₃₋₆cycloalkyl,C₃₋₆alkenyl, hydroxyC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl orC₁₋₆alkyloxyC₁₋₆alkyl; R⁵ is C₁₋₈alkyl, mono- ordi(C₃₋₆cycloalkyl)methyl, Ar¹CH₂, C₃₋₆alkenyl, C₁₋₆alkyloxyC₁₋₆alkyl,hydroxyC₁₋₆alkyl, thienylmethyl, furanylmethyl, C₁₋₆alkylthioC₁₋₆alkyl,morpholinyl, mono- or di(C₁₋₆alkyl)aminoC₁₋₆alkyl, di(C₁₋₆alkyl)amino,C₁₋₆alkylcarbonylC₁₋₆alkyl, C₁₋₆alkyl substituted with imidazolyl; or aradical of formula —Alk—O—CO—Ar¹; or R⁴ and R⁵ taken together with thenitrogen atom to which they are attached may form a pyrrolidinyl,piperidinyl, homopiperidinyl or morpholinyl group, optionallysubstituted with C₁₋₆alkyl or C₁₋₆alkyloxyC₁₋₆alkyl; and Ar is phenyl;phenyl substituted with 1, 2 or 3 substituents independently selectedfrom halo, C₁₋₆alkyl, trifluoromethyl, hydroxy, cyano, C₁₋₆alkyloxy,benzyloxy, C₁₋₆alkylthio, nitro, amino and mono- or di(C₁₋₆alkyl)amino;pyridinyl; pyridinyl substituted with 1, 2 or 3 substituentsindependently selected from halo, C₁₋₆alkyl, trifluoromethyl, hydroxy,cyano, C₁₋₆alkyloxy, benzyloxy, C₁₋₆alkylthio, nitro, amino, mono- ordi(C₁₋₆alkyl)amino and piperidinyl; and wherein said substituted phenylmay optionally be further substituted with one or more halogens; Ar¹ isphenyl; phenyl substituted with 1, 2 or 3 substituents eachindependently selected from halo, C₁₋₆alkyl, C₁₋₆alkyloxy,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, trifluoromethyl and C₁₋₆alkyl substitutedwith morpholinyl; or pyridinyl; and Alk is C₁₋₆alkanediyl; with theproviso that5-methyl-3-phenyl-7-(phenylmethoxy)-pyrazolo[1,5-a]-pyrimidine and2,5-dimethyl-7-(methylamino)-3-phenyl-pyrazolo[1,5-a]pyrimidine are notincluded.
 2. A compound according to claim 1 wherein R¹ is OR⁵ and R⁵ isC₁₋₆alkyl; or R¹ is NR⁴R⁵ and R⁴ is hydrogen, C₁₋₆alkyl,hydroxyC₁₋₆alkyl, C₁₋₆alkylcarbonyloxy-C₁₋₆alkyl or C₃₋₆alkenyl, R⁵ isC₁₋₈alkyl, C₃₋₆alkenyl, hydroxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl,phenylmethyl or C₃₋₆cycloalkylmethyl; or R⁴ and R⁵ are taken togetherwith the nitrogen atom to which they are attached to form apyrrolidinyl, piperidinyl, homopiperidinyl or morpholinyl group,optionally substituted with C₁₋₆alkyl or C₁₋₆alkyloxyC₁₋₆alkyl; R² isC₁₋₆alkyl; R³ is hydrogen, C₁₋₆alkyl or C₁₋₆alkylthio; and Ar is aphenyl substituted with 1, 2 or 3 substituents each independentlyselected from halo, C₁₋₆alkyl or C₁₋₆alkyloxy; or Ar is a pyridinylsubstituted with 1, 2 or 3 substituents each independently selected fromhalo, amino, nitro, trifluoromethyl, mono- or di(C₁₋₆alkyl)amino,piperidinyl or C₁₋₆alkyl.
 3. A compound according to any of claims 1 to2 wherein R¹ is NR⁴R⁵ and R⁴ is C₂₋₄alkyl, hydroxyC₁₋₂alkyl, C₃₋₄alkenylor C₁₋₂alkylcarbonyloxyC₂₋₄alkyl; R⁵ is C₂₋₄alkyl, C₃₋₄alkenyl,hydroxyC₂₋₄alkyl or cyclopropylmethyl; R² is C₁₋₂alkyl; R³ is hydrogen,C₁₋₂alkyl or C₁₋₂alkylthio.
 4. A compound according to any of claims 1to 3 wherein R¹ is NR⁴R⁵ and R⁴ is C₃₋₄alkyl or allyl; R⁵ is C₂₋₄alkyl,allyl or cyclopropylmethyl; R² is methyl; R³ is methyl; and Ar is phenylsubstituted in the 3-, 4-, 6-, 2,4- or 2,4,6-positions with halo,C₁₋₆alkyl or C₁₋₆alkyloxy.
 5. A compound according to any of claims 1 to3 wherein R¹ is NR⁴R⁵ and R⁴ is C₃₋₄alkyl or allyl; R⁵ is C₃₋₄alkyl,allyl or cyclopropylmethyl; R² is methyl; R³ is methyl; and Ar is3-pyridinyl substituted on the 4- and/or 6-position with methyl ordimethylamino.
 6. A compound according to claim 1 wherein the compoundis3-[6-(dimethylamino)-3-pyridinyl]-2,5-dimethyl-N,N-dipropylpyrazolo[2,3-a]pyrimidin-7-amine,or3-[6-(dimethylamino)-4-methyl-3-pyridinyl]-2,5-dimethyl-N,N-dipropylpyrazolo[2,3-a]pyrimidin-7-amine;the stereochemically isomeric forms, or the pharmaceutically acceptableacid addition salts thereof.
 7. A composition comprising apharmaceutically acceptable carrier, and as active ingredient atherapeutically effective amount of a compound as claimed in any one ofclaims 1 to
 6. 8. A process for preparing a composition as claimed inclaim 7 characterized in that a therapeutically effective amount of acompound as claimed in any one of claims 1 to 6 is intimately mixed witha pharmaceutically acceptable carrier.
 9. A compound according to anyone of claims 1 to 6 for use as a medicine.
 10. The use of compounds offormula

including the stereoisomers and the pharmaceutically acceptable acidaddition salt forms thereof, wherein R¹, R², R³ and Ar are as defined inclaim 1, including the compounds5-methyl-3-phenyl-7-(phenylmethoxy)-pyrazolo[1,5-a]-pyrimidine and2,5-dimethyl-7-(methylamino)-3-phenyl-pyrazolo[1,5-a]pyrimidine, for themanufacture of a medicament for treating physiological conditions ordisorders arising from the hypersecretion of corticotropin-releasingfactor (CRF).
 11. A compound of formula (II′) wherein the radicals R²,R³ and Ar are as defined in any of claims 1 to 5 and radical W′ ishydroxy, halo, mesyloxy or tosyloxy; a stereoisomeric form or an acidaddition salt form thereof, with the proviso that compounds wherein Aris unsubstituted phenyl are not included.


12. A process of preparing a compound as claimed in claim 1,characterized by a) reacting an intermediate of formula (II) with anintermediate of formula (III) in a reaction-inert solvent,

b) O-alkylating an intermediate of formula (VI) with an intermediate offormula (VII) in a reaction-inert solvent and in the presence of asuitable base, yielding compounds of formula (I-a), defined as compoundsof formula (I) wherein R¹ is OR⁵,

wherein in the above reaction schemes the radicals R¹, R², R³, R⁵ and Arare as defined in claim 1 and W is an appropriate leaving group; or, ifdesired, converting compounds of formula (I) into each other followingart-known transformation reactions; and further, if desired, convertingthe compounds of formula (I), into an acid addition salt by treatmentwith an acid, or conversely, converting the acid addition salt form intothe free base by treatment with alkali; and, if desired, preparingstereochemically isomeric forms thereof.
 13. A process of preparing acompound of formula (II′) as claimed in claim 10 characterized byreacting an intermediate of formula (IV) with a β-keto ester (V) in areaction-inert solvent, thereby yielding compounds of formula (II′-a),defined as compounds of formula (II′) wherein W′ is hydroxy;

and optionally converting compounds of formula (II′-a) into compounds offormula (II′-b), defined as compounds of formula (II′) wherein W′ isother than hydroxy; wherein in the above reaction schemes the radicalsR², R³ and Ar are as defined in claim 1 and W′ is hydroxy, halo,mesyloxy or tosyloxy; or, if desired, converting compounds of formula(II′) into each other following art-known transformation reactions; andfurther, if desired, converting the compounds of formula (II′), into anacid addition salt by treatment with an acid, or conversely, convertingthe acid addition salt form into the free base by treatment with alkali;and, if desired, preparing stereochemically isomeric forms thereof.